摘要
目的:探讨雌激素受体(ER)与表皮生长因子受体(EGFR)的关系及其在乳腺癌三苯氧胺(TAM)治疗耐药中的作用与机制。方法:选用TAM治疗敏感(TAM-S)与耐药(TAM-R)的两种人乳腺癌MCF-7细胞,用免疫沉淀(IP)法检测ERα和EGFR在两种细胞中的结合情况,以及c-Src与前两者的结合情况,并用Western blot法检测两种细胞中c-Src的磷酸化水平;c-Src阻滞剂PP2处理两种细胞后,IP法再次检测ERα和EGFR结合情况。结果:ERα和EGFR在两种细胞中均以复合物的形式结合,但在TAM-R细胞中的结合量明显高于TAM-S细胞(P<0.05);c-Src与ERα、EGFR在两种细胞中均以复合物的形式结合,其磷酸化水平在TAM-R细胞中明显高于TAM-S细胞(P<0.05);PP2阻断c-Src的活性后,两种细胞中ERα和EGFR结合量均降低,且在TAM-R细胞中的降低程度明显大于TAM-S细胞(P<0.05)。结论:ERα和EGFR之间存在结合的现象,且两者的互作关系可能在乳腺癌TAM治疗耐药中起了重要作用,而c-Src的活化(磷酸化)是可能是介导了两者结合的关键机制。
Objective: To investigate the relationship between estrogen receptor (ER) and epidermal growth factor receptor (EGFR), and its role and mechanism in tamoxifen (TAM) therapy resistance of breast cancer.Methods: Two types of cells namely the TAM therapy-sensitive (TAM-S) and -resistant (TAM-R) human breast cancer MCF-7 cells were used. In these two types of cells, immunoprecipitation (IP) was performed to detect the connection between ERα and EGFR, and the connection of c-Src to the two former receptors, and the phosphorylation level of c-Src was also determined by Western blot analysis. After treatment with c-Src inhibitor PP2, the connection between ERα and EGFR was examined again by IP method. Results: ERα and EGFR linked in a complex form in both types of cells, but the linkage level in TAM-R cells was significantly higher than that in TAM-S cells (P〈0.05). c-Src linked in a complex form to either ERα or EGFR in both types of cells, and the phosphorylation level of c-Src in TAM-R cells was significantly higher than that in TAM-S cells (P〈0.05). After the c-Src activity was inhibited by PP2, linkage level between ERα or EGFR was decreased in both types of cells, while the decreasing degree in TAM-R cells was significantly greater than that in TAM-S cells (P〈0.05).Conclusion: There is a connection between ERα and EGFR, which may play an important role in TAM therapy resistance of breast cancer, and c-Src activation (phosphorylation) may be the crucial mechanism for their linkage.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2014年第5期618-623,共6页
China Journal of General Surgery
基金
江苏省无锡市卫生局科研资助项目(XM1010)